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MICROBIOLOGY
E2 E1 Core RNA
Cytoplasm
E1 or E2 receptor binding
Target
Viral core release Polymerase inhibitors 3 NS5B Endoplasmic reticulum NS3 Helicase inhibitors 5 Viral release
Core (191 amino acids) IRES (340 bases)
Function
Initiation of cap-independent translation Viral nucleocapsid and RNA packaging
Attractive features
Highly conserved and structured
Challenges
Specificity of non-oligonucleotide inhibitors has not been demonstrated Protein is poorly structured prior to assembly; corecore interaction avidity can reduce potency of any inhibitors in vivo Low sequence conservation
Endocytosis
Fusion
Processing NS3 NS3 protease inhibitors Viral budding and particle formation
Cysteine autoprotease
Non-structural protein
Development of autoproteolysis assays is challenging; small molecule inhibition of activity not shown No structural information known, except that the central portion binds to NS3. Direct binding of ligands has not been demonstrated
Nucleus
References
1. 2. 3. 4. von Hahn, T. & Rice, C. M. Hepatitis C virus entry. J. Biol. Chem. 283, 36893693 (2008). Ploss, A. et al. Human occludin is a hepatitis C virus entry factor required for infection of mouse cells. Nature 457, 882886 (2009). Soriano, V., Peters, M. G. & Zeuzem, S. New therapies for hepatitis C virus infection. Clin. Infect. Dis. 48, 313320 (2009). Kwong, A. D., McNair, L., Jacobson, I. & George, S. Recent progress in the development of selected 5.
6.
hepatitis C virus NS3.4A protease and NS5B polymerase inhibitors. Curr. Opin. Pharmacol. 8, 522531 (2008). Beaulieu, P. L. Recent advances in the development of NS5B polymerase inhibitors for the treatment of hepatitis C virus infection. Expert Opin. Ther. Patents 19, 145164 (2009). Huang, H. et al. Hepatitis C virus production by human hepatocytes dependent on assembly and secretion of very low-density
7.
8.
lipoproteins. Proc. Natl Acad. Sci. USA 104, 58485853 (2007). Gastaminza, P. et al. Cellular determinants of hepatitis C virus assembly, maturation, degradation, and secretion. J. Virol. 82, 21202129 (2008). Jones, C. T., Murray, C. L., Eastman, D. K., Tassello, J. & Rice, C. M. Hepatitis C virus p7 and NS2 proteins are essential for infectious virus production. J. Virol. 81, 83748383 (2007).
Ma, Y., Yates, J., Liang, Y., Lemon, S. M. & Yi, M. K. NS3 helicase domains involved in infectious intracellular hepatitis C virus particle assembly. J. Virol. 82, 76247639 (2008). 10. Flisiak, R. et al. The cyclophilin inhibitor Debio-025 shows potent anti-hepatitis C effect in patients coinfected with hepatitis C and human immunodeficiency virus. Hepatology 47, 817826 (2008).
9.
Affiliation
Richard Bethell, George Kukolj and Peter W. White are at Boehringer Ingelheim (Canada), 2100, rue Cunard, Laval (Qubec), H7S 2G5, Canada.
Acknowledgements
The authors acknowledge R. Coulombe for the preparation of the protein structure images used in this poster. The content of this poster has been peerreviewed, similarly to all articles published in Nature Reviews Microbiology.
Edited by Christiaan van Ooij; copy-edited by Gillian Young; designed by George Marshall. 2009 Nature Publishing Group. For further reading, see http://www.nature.com/ nrmicro/posters/hepatitis-c/
C-terminal region not well structured in vitro; direct binding of ligands to protein has not been demonstrated
SUPPLEMENTARY INFORMATION
Supplementary information S1 | Further reading
Virology
Choo Q. L., et al. Isolation of a cDNA clone derived from a blood-borne non-A, non-B viral hepatitis genome. Science 244, 359362 (1989). Lohmann V., et al. Replication of subgenomic hepatitis C virus RNAs in a hepatoma cell line. Science 285, 110113 (1999). Wakita, T. et al. Production of infectious hepatitis C virus in tissue culture from a cloned viral genome. Nature Med. 11, 791796 (2005). Lindenbach B. D., et al. Complete replication of hepatitis C virus in cell culture. Science 309, 623626 (2005). Moaradpour, D., Penin, F. & Rice, C. M. Replication of hepatitis C virus Nature Rev. Microbiol. 5, 453463 (2007).
Core
Murray, C. L., Jones, C. T., Tassello, J. & Rice, C. M. Alanine scanning of the hepatitis C virus core protein reveals numerous residues essential for production of infectious virus J. Virol. 81, 1022010231 (2007). Hourioux, C. et al. Core protein domains involved in hepatitis C virus-like particle assembly and budding at the endoplasmic reticulum membrane Cell. Microbiol. 9, 10141027 (2007). Ivanyi-Naggy, R., Lavergne, J.-P., Gabus, C., Ficheux, D., Darlix, J.-L. RNA chaperoning and intrinsic disorder in the core proteins of Flaviviridae Nucl. Acids Res. 36, 712725 (2008).
NS4A Yang, W., et al. Selection of replicon variants resistant to ACH-806, a novel hepatitis C virus inhibitor with no cross-resistance to NS3 protease and NS5B polymerase inhibitors Antimicrob. Agents Chemother. 52, 20432052 (2008).
NS4B
Jones, D. M., Patel, A. H., Targett-Adams, P. & McLauchlan, J. The hepatitis C virus NS4B protein can trans-complement viral RNA replication and modulates production of infectious virus J. Virol. 83, 21632177 (2009). Thompson, A. A., et al. Biochemical characterization of recombinant hepatitis C virus non-structural protein 4B: evidence for ATP/GTP hydrolysis and adenylate kinase activity Biochemistry 48, 909916 (2009). Einav, S., et al. Discovery of a hepatitis C target and its pharmacological inhibitors by microfluidic affinity analysis Nature Biotech. 26, 10191027 (2008).
p7
Steinmann, E. et al. Hepatitis C virus p7 protein is crucial for assembly and release of infectious virions PLoS Path. 3, 962971 (2007). Griffin, S. et al. Genotype-dependent sensitivity of hepatitis C virus to inhibitors of the p7 ion channel Hepatology 48, 17791790 (2008).
Drug Discovery
Pawlotsky, J.-M., Chevaliez S., & McHutchison, J. G. The hepatitis C virus life cycle as a target for new antiviral therapies Gastroenterol. 132, 19791998 (2007). Asselah, R., Benhamou, Y. & Marcellin, P. Protease and polymerase inhibitors for the treatment of hepatitis C Liver International 29, 5767 (2009).
NS5A
NS2
Welbourn, S. et al. Hepatits C virus NS2/3 processing is required for NS3 stability and viral RNA replication J. Biol. Chem. 280, 2960429611 (2005). Schregel, V., Jacobi, S., Penin, F. & Tautz, N. Hepatitis C virus NS2 is a protease stimulated by cofactor domains in NS3 Proc. Natl Acad. Sci. USA 106, 53425347 (2009). Lorenz, I. C., Marcotrigiano, J., Dentzer, T. G. & Rice, C. M. Structure of the catalytic domain of the hepatitis C virus NS23 protease Nature 442, 831835 (2006).
Macdonald A. & Harris, M. Hepatitis C virus NS5A: tales of a promiscuous protein J. Gen.Virol. 85, 2485 2502 (2004). Tellinghuisen, T. L., Foss, K. L. & Treadaway, J. Regulation of hepatitis C virion production via phosphorylation of the NS5A protein PLoS Path. 4(3), e1000032 (2008) Appel, N., et al. Essential role of domain III nonstructutal protein 5A for hepatitis C virus infectious particle assembly Plos Path. 4(3), e1000035 (2008). Tellinghuisen, T. L., Marcotrigiano, J. & Rice, C. M. Structure of the zinc-binding domain of an essential component of the hepatitis C virus replicase Nature 435, 374379 (2005). Love, R. A., Brodsky, O., Hickey, M. J., Wells, P. A. & Cronin, C. N. Crystal structure of a novel dimeric form of NS5A domain I protein from hepatitis C virus J. Virol. 83, 4395-4403 (2009). Schmitz, U. & Tan, S.-L. NS5A- from obscurity to new target for HCV therapy Recent Pat. Anti-Inf. Drug Disc. 3, 7792 (2008). Nettles, R., et al. BMS-790052 is a first -in-class potent hepatitis C virus (HCV) NS5A inhibitor for patients with chronic HCV infection: results from a proof-of-concept study AASLD Annual Meeting LB12 (2008).
Entry
Pileri P. et al. Binding of hepatitis C virus to CD81. Science 282, 938941 (1998). Scarselli, E. et al. The human scavenger receptor class B type I is a novel candidate receptor for the hepatitis C virus. EMBO J. 21, 50175025 (2002). Lozach P.Y. et al. DC-SIGN and L-SIGN are high affinity binding receptors for hepatitis C virus glycoprotein E2. J. Biol. Chem. 278, 2035820366 (2003). Cormier E.G. et al. L-SIGN (CD209L) and DC-SIGN (CD209) mediate transinfection of liver cells by hepatitis C virus. Proc. Natl Acad. Sci. USA. 101, 1406772 (2004). Evans, M.J. et al. Claudin-1 is a hepatitis C virus co-receptor required for a late step in entry. Nature 446, 801805 (2007). Dubuisson, J., Helle F. & Cocquerel L. Early steps of the hepatitis C virus life cycle. Cell. Microbiol. 10, 821 827 (2008).
NS3 protease
Kim, J. L., et al. Crystal structure of the hepatitis C virus NS3 protease domain complexed with a synthetic NS4A cofactor peptide Cell 87, 343355 (1996). Yao, N., Reichert, P., Taremi, S. S., Prosise, W. W. & Weber, P. C. Molecular views of viral polyprotein processing revealed by the crystal structure of the hepatitis C virus bifunctional protease-helicase Structure 7, 13531363 (1999). Lamarre, D., et al. An NS3 protease inhibitor with antiviral effects in humans infected with hepatitis C virus Nature 426, 186189 (2003). Hinrichsen, H., et al. Short-term antiviral efficacy of BILN 2061, a hepatitis C virus serine protease inhibitor, in hepatitis C genotype 1 patients Gastroenterology 127, 13471355 (2004). Gentile, I., Viola, C., Borgia, F., Castaldo, G. & Borgia, G. Telaprevir: a promising protease inhibitor for the treatment of hepatitits C virus infection Curr. Med. Chem. 16, 11151121 (2009). Njoroge, F. G., Chen, K. X., Shih, N.-Y. & Piwinski, J. J. Challenges in modern drug discovery: a case study of boceprevir, an HCV protease inhibitor for the treatment of hepatitis C virus infection Accts. Chem. Res. 41, 5059 (2008).
Assembly
Miyanari Y., et al. The lipid droplet is an important organelle for hepatitis C virus production. Nature Cell Biol. 9, 10891097 (2007). Roingeard P, Hourioux C, Blanchard E & Prensier G. Hepatitis C virus budding at lipid droplet-associated ER membrane visualized by 3D electron microscopy. Histochem. Cell Biol. 130, 561566 (2008).
NS5B
Behrens SE, Tomei L & De Francesco R. Identification and properties of the RNA-dependent RNA polymerase of hepatitis C virus. EMBO J. 15, 1222 (1996). Lohmann V, Krner F, Herian U & Bartenschlager R. Biochemical properties of hepatitis C virus NS5B RNAdependent RNA polymerase and identification of amino acid sequence motifs essential for enzymatic activity. J Virol. 71, 84168428 (1997). De Francesco R, Carf A. Advances in the development of new therapeutic agents targeting the NS34A serine protease or the NS5B RNA-dependent RNA polymerase of the hepatitis C virus. Adv Drug Deliv Rev. 59, 12421262 (2007).
Targets Host
Cyclophilin
Crabb, R. et al. An evaluation of the cyclophilin inhibitor Debio 025 and its potential as a treatment for chronic hepatitis C Expert Opin. Investig. Drugs 18, 211220 (2009).
Targets Viral
IRES
Lukavsky, P. J. (2009). Structure and function of HCV IRES domains Virus Res. 139, 166171 McHutchison, J. G. et al. A phase I trial of an antisense inhibitor of hepatitis C virus (ISIS 14803), administered to chronic hepatitis C patients J. Hepatol. 44, 8896 (2006).
NS3 helicase
Frick, D. N. The hepatitis C virus NS3 protein: a model RNA helicase and potential drug target Curr. Issues Mol. Biol. 9, 120 (2007). Mackintosh, S. G.,et al. Structural and biological identification of residues on the surface of NS3 helicase required for optimal replication of the hepatitis C virus J. Biol. Chem. 281, 35283535 (2006).
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